降解纤维素凝结芽孢杆菌的分离鉴定及生物学特性研究  

Isolation,Identification and Biological Characteristic of Acellulase-producing Bacillus coagulans

作　　者：余祖玲 余祖华 贾艳艳 廖成水 李旺[1] 李元晓[1] 何万领[1] 丁轲[1,2] YU Zuling;YU Zuhua;JIA Yanyan;LIAO Chengshui;LI Wang;LI Yuanxiao;HE Wanling;DING Ke(Functional Microbiology and Precision Nutrition Laboratory,Henan University of Science&Technology,Luoyang 471023,China;Luoyang Key Laboratory of Live Carrier Biomaterials and Animal Disease Prevention and Control,Luoyang 471023,China)

机构地区：[1]河南科技大学功能微生物与精准营养实验室,河南洛阳471023 [2]洛阳市活载体生物材料与动物疫病防控重点实验室,河南洛阳471023

出　　处：《河南科技大学学报（自然科学版）》2022年第5期84-91,99,M0007,M0008,共11页Journal of Henan University of Science And Technology:Natural Science

基　　金：国家自然科学基金项目(32072771);河南省自然科学基金项目(182300410052)。

摘　　要：为筛选降解纤维素性能优良的芽孢杆菌,探究其生物学特性,采集不同地方的土壤作为样品,将其接种到羧甲基纤维素钠(CMC-Na)培养基进行分离。观察菌株是否产生水解圈进行初筛,再通过检测滤纸酶活和纤维素酶活进行复筛。通过形态学观察、生理生化及16S rDNA测序对菌株进行鉴定,并对分离菌株进行胆盐、酸、人工胃液、高温的耐受能力测试及药敏试验。筛选获得了1株降解纤维素功能较强的菌株MRS-913,经鉴定为凝结芽孢杆菌(Bacillus coagulans)。生物学性能试验表明:该菌株在100℃水浴中15 min存活率达54.0%;在质量分数为4%的胆盐中4 h存活率为32.8%;在pH1.0的酸液中4 h存活率为55.9%;在pH1.5和pH2.5的模拟胃液中3 h存活率分别为62.7%和74.8%。药敏试验结果表明:该菌株对青霉素、氟苯尼考和头孢曲松高度敏感,对杆菌肽、奥普托欣和林可霉素不敏感。因此,凝结芽孢杆菌MRS-913有望成为微生态制剂的候选菌株。In order to isolate and screen out bacillus with good performance of degrading cellulose,the biological characteristics was explored.In this paper,soil samples from different places were collected as samples and inoculated into sodium carboxymethyl cellulose(CMC-NA)medium for separation.Primary screening was conducted by observing whether the strain produced hydrolytic circles or not,and then rescreening was conducted by detecting the enzyme activity of filter paper and cellulase activity.The strains were identified by morphological observation,physiological and biochemical analysis and 16SrDNA sequencing.The strains were tested for tolerance of bile salt,acid,artificial gastric juice and high temperature.A cellulose-degrading strain MRS-913 was screened and identified as Bacillus coagulans.The biological test shows that the survival rate of the strain is 54.0%after 15 min of 100℃water bath.The survival rate for 4 h in 4%bile salt is 32.8%.The survival rate of 4 h in pH 1.0 acid is 55.9%.The 3 h survival rate in simulated gastric juice at pH1.5 and pH2.5 is 62.7%and 74.8%,respectively.The results of antimicrobial susceptibility test show that the strain is highly sensitive to penicillin,flufenicol and ceftriaxone,but not to bacitracin,optophin and lincomycin.Therefore,bacillus coagulans MRS-913 is expected to be developed as a candidate strain of probiotics.

关 键 词：纤维素 凝结芽孢杆菌 分离鉴定 生物学特性 

分 类 号：S816.7[农业科学—饲料科学]